2.1 Prepare transcription solution
Prepare the transcription reaction solution in a 1.5 ml tube as follows.
Do not mix 2nd PCR Product directly with 10x Transcription Buffer.
| Reagents | Volume |
| 10x Transcription Buffer | 2.5 μl |
| 25 mM NTP Mix | 2.5 μl |
| T7 RNA Polymerase | 1 μl |
| 100 mM DTT | 1.25 μl |
| 2nd PCR Product | 2.5 μl |
| RNase free water | 15.25 μl |
| total | 25 μl |